Clinically applied procedures for human ovarian tissue cryopreservation result in different levels of efficacy and efficiency

Purpose

Different protocols are being used worldwide for the cryopreservation of human ovarian tissue for fertility preservation purposes. The efficiency and efficacy of the majority of these protocols has not been extensively evaluated, possibly resulting in sub-optimally cryopreserved ovarian tissue. To address the impact of this issue, we assessed the effects of two clinically successful human ovarian tissue slow-freezing cryopreservation procedures on the quality of the cryopreserved tissue.

Methods

To differentiate between cryopreservation (_C) versus thawing (_T) related effects, four combinations of these two (A and B) very different cryopreservation/thawing protocols (A_CA_T, A_CB_T, B_CA_T, B_CB_T) were studied. Before and after cryopreservation and thawing, the percentage of living and morphologically normal follicles, as well as the overall tissue viability, was assessed.

Results

Our experiments revealed that the choice of the cryopreservation protocol noticeably affected the overall tissue viability and percentage of living follicles, with a higher viability after protocol B_C when compared to A_C. No statistically significant differences in tissue viability were observed between the two thawing protocols, but thawing protocol B_T required considerably more human effort and materials than thawing protocol A_T. Tissue morphology was best retained using the B_CA_T combination.

Conclusion

Our results indicate that extensive and systematical evaluation of clinically used protocols is warranted.

     

A systematic description of the post-EMR defect to identify risk factors for clinically significant post-EMR bleeding in the colon

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Tumor suppressor Foxo3a is involved in the regulation of lipopolysaccharide-induced interleukin-8 in intestinal HT-29 cells

Enteric bacteria and their products play an important role in intestinal inflammation; however, the complete mechanisms are not elucidated yet. Tumor suppressor Foxo3a regulates gene expression in the nucleus, and its translocation to the cytosol leads to inactivation. Proximally, Foxo3a is regulated by different pathways including the phosphoinositide 3-kinase (PI3K) pathway. The aim of this study was to determine the effect of bacterial infection on Foxo3a in intestinal epithelial cells and to examine the contribution of Foxo3a in intestinal inflammation. Bacterial lipopolysaccharide (LPS) and infection with mouse pathogen Citrobacter rodentium induce translocation of the nuclear Foxo3a into the cytosol, where it degrades in human HT-29 and mouse CMT-93 cells. In colonic epithelia of healthy mice, Foxo3a is localized in the epithelia at the bottom of the crypts in both the nucleus and the cytosol, while in C. rodentium-infected colon Foxo3a is expressed along the crypts and located mainly in the cytosol, suggesting its inactivation. LPS utilized the PI3K pathway to inhibit Foxo3a. Additionally, inhibition of PI3K attenuated LPS-induced proinflammatory interleukin-8 (IL-8). LPS-induced IL-8 is increased in HT-29 cells with silenced Foxo3a. Moreover, in HT-29 cells with silenced Foxo3a, the amount of IκBα, an NF-κB inhibitor, is decreased. In conclusion, LPS and bacterial infection inactivate Foxo3a in intestinal epithelia via the PI3K pathway and inactivated Foxo3a leads to the upregulation of IL-8 by suppressing inhibitory IκBα.     

Evaluation of a psycho‐educational group intervention for children treated for cancer: a descriptive pilot study

Objective: The present paper reports about the content and evaluation of a psycho‐educational group intervention for children growing up with a history of cancer, Op Koers Oncologie (OK Onco). OK Onco is aimed at empowerment of survivors of childhood cancer by teaching disease‐related skills. The purpose of this pilot study is to evaluate whether OK Onco was appropriate for use among paediatric survivors of childhood cancer. Methods: Eleven participating children and their parents completed questionnaires about the disease‐related skills that were taught in the intervention, before the intervention and 0–4 weeks after the intervention. In addition, parents were interviewed in focus groups. Results: Positive outcomes were found on most items concerning disease‐related skills. Remarkable improvements were found with respect to the intervention goals ‘social competence’ and ‘positive thinking’. The parents reported that sharing emotions and experiences with other survivors of childhood cancer was an important element of the OK Onco program, which underscores the benefit of a disease‐specific program for cancer survivors. Conclusions: In conclusion, the results of the pilot study indicate that OK Onco is appropriate for paediatric survivors of childhood cancer. Implementation of the intervention is the next challenge. Copyright © 2008 John Wiley & Sons, Ltd.     

A Study of Carry-Over and Histopathological Effects after Chronic Dietary Intake of Citrinin in Pigs,Broiler Chickens and Laying Hens

Citrinin (CIT) is a polyketide mycotoxin occurring in a variety of food and feedstuff, among which cereal grains are the most important contaminated source. Pigs and poultry are important livestock animals frequently exposed to mycotoxins, including CIT. Concerns are rising related to the toxic, and especially the potential nephrotoxic, properties of CIT. The purpose of this study was to clarify the histopathological effects on kidneys, liver, jejunum and duodenum of pigs, broiler chickens and laying hens receiving CIT contaminated feed. During 3 weeks, pigs (n = 16) were exposed to feed containing 1 mg CIT/kg feed or to control feed (n = 4), while 2 groups of broiler chickens and laying hens (n = 8 per group) received 0.1 mg CIT/kg feed (lower dose group) and 3 or 3.5 mg CIT/kg feed (higher dose group), respectively, or control feed (n = 4). CIT concentrations were quantified in plasma, kidneys, liver, muscle and eggs using a validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. Kidneys, liver, duodenum and jejunum were evaluated histologically using light microscopy, while the kidneys were further examined using transmission electron microscopy (TEM). Histopathology did not reveal major abnormalities at the given contamination levels. However, a significant increase of swollen and degenerated mitochondria in renal cortical cells from all test groups were observed (p < 0.05). These observations could be related to oxidative stress, which is the major mechanism of CIT toxicity. Residues of CIT were detected in all collected tissues, except for muscle and egg white from layers in the lowest dose group, and egg white from layers in the highest dose group. CIT concentrations in plasma ranged between 0.1 (laying hens in lower dose group) and 20.8 ng/mL (pigs). In tissues, CIT concentrations ranged from 0.6 (muscle) to 20.3 µg/kg (liver) in pigs, while concentrations in chickens ranged from 0.1 (muscle) to 70.2 µg/kg (liver). Carry-over ratios from feed to edible tissues were between 0.1 and 2% in pigs, and between 0.1 and 6.9% in chickens, suggesting a low contribution of pig and poultry tissue-derived products towards the total dietary CIT intake for humans.     

Sensitivity of Spontaneous Baroreflex Control of the Heart and Hemodynamic Parameters are Not Influenced by the Menstrual Cycle

Objective: The purpose of this study was to evaluate if hemodynamic parameters and sympathetic activity vary between the follicular and luteal phase of the menstrual cycle before using sympathetic activity in pre-pregnancy risk assessment for preeclampsia. Methods: We studied 39 healthy women at days 5 to 10 and days 18 to 25 of the menstrual cycle. Blood pressure, heart rate, cardiac output, and total peripheral resistance were measured continuously using noninvasive finger arterial pressure waveform registration (Portapres Model 2, BMI, The Netherlands). Baroreflex sensitivity (BRS) and sympathetic activity by phase angle difference were studied using spectral analysis and xBRS. Results: There were no differences in hemodynamic parameters, BRS, and phase angle difference. Conclusion: There is no difference in blood pressure, BRS, and sympathetic activity between the first and second half of the menstrual period. We recommend using the first half of the cycle to be certain that no pregnancy exists, as the influence of very early pregnancy is unknown.     

High live birth rate in the subsequent IVF cycle after first-cycle poor response among women with mean age 35 and normal FSH

Poor ovarian response in IVF cycles is associated with diminished ovarian reserve and poor pregnancy outcome. Little is known about pregnancy outcome after a poor response in women with a normal ovarian reserve. This retrospective study studied women undergoing IVF/intracytoplasmic sperm injection between January 2003 to December 2008 in the FertilityPLUS Clinic in Auckland, New Zealand. All women with a poor response in the first cycle were selected. Primary outcome was live birth after the second cycle. Secondary outcomes were poor response in the second cycle and the predictive values of female age and basal FSH at first cycle and IVF outcome at second cycle. Of the 2487 women starting IVF, 142 women (5.7%) with a poor response in the first cycle were selected, of which 66 (46.5%) women had a repeated poor response in the second cycle. There were 31 live births in the second cycle (21.8%). Female age was the only significant predictor for repeated poor response (AUC 0.69, 95% CI 0.61–0.78) and clinical pregnancy (AUC 0.66, 95% CI 0.57–0.75), but the predictive value was low. Therefore poor response in women with a normal ovarian reserve should not be a reason to discontinue further IVF treatment.Poor ovarian response in IVF cycles is associated with diminished ovarian reserve and poor pregnancy outcome. Little is known about pregnancy outcome after a poor response in women with a normal ovarian reserve. In this retrospective study, we studied women undergoing IVF/intracytoplasmic sperm injection (ICSI) between January 2003 to December 2008 in the FertilityPLUS Clinic in Auckland, New Zealand. All women with a poor response in the first cycle were selected. Primary outcome was live birth after the second cycle. Secondary outcomes were poor response in the second cycle and the predictive value of female age and basal FSH at first cycle and IVF outcome at the second cycle. Of the 2487 women starting wit IVF, a total of 142 women (5.7%) with a poor response in the first cycle were selected, of which 66 (46.5%) women had a repeated poor response in the second cycle. There were 31 live births in the second cycle (22%). Female age was the only significant predictor for repeated poor response and clinical pregnancy, but the predictive value was low. Therefore poor response in women with a normal ovarian reserve should not be a reason to discontinue further IVF treatment.